Nanoparticles having oligonucleotides attached thereto and uses therefor

a technology of oligonucleotides and nanoparticles, which is applied in the field of nanoparticles having oligonucleotides attached thereto and uses therefor, can solve the problems of complex methods, time-consuming and/or expensive equipment, and assembly formation irreversibly,

Inactive Publication Date: 2002-10-24
NORTHWESTERN UNIV +6
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

0007] The invention provides methods of detecting nucleic acids. In one embodiment, the method comprises contacting a nucleic acid with a type of nanoparticles having oligonucleotides attached thereto (nanoparticle-oligonucleotide conjugates). The nucleic acid has at least two portions, and the oligonucleotides on each nanoparticle have a sequence complementary to the sequences of at least two portions of the nucleic acid. The contacting takes place under conditions effective to allow hybridization of the oligonucleotides on the nanoparticles with the nucleic acid. The hybridization of the oligonucleotides on the nanoparticles with the nucleic acid results in a detectable change.

Problems solved by technology

However, these methods are complicated, time-consuming and / or require the use of specialized and expensive equipment.
The drawbacks of this method are that the process is difficult to control and the assemblies are formed irreversibly.
However, the theory of producing DNA structures is well ahead of experimental confirmation.

Method used

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  • Nanoparticles having oligonucleotides attached thereto and uses therefor
  • Nanoparticles having oligonucleotides attached thereto and uses therefor
  • Nanoparticles having oligonucleotides attached thereto and uses therefor

Examples

Experimental program
Comparison scheme
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second embodiment

[0214] In a second embodiment, the kit comprises at least two containers. The first container holds nanoparticles having oligonucleotides attached thereto which have a sequence complementary to the sequence of a first portion of a nucleic acid. The second container holds nanoparticles having oligonucleotides attached thereto which have a sequence complementary to the sequence of a second portion of the nucleic acid. The kit may further comprise a third container holding a filler oligonucleotide having a sequence complementary to a third portion of the nucleic acid, the third portion being located between the first and second portions.

[0215] In another alternative embodiment, the kits can have the oligonucleotides and nanoparticles in separate containers, and the oligonucleotides would have to be attached to the nanoparticles prior to performing an assay to detect a nucleic acid. The oligonucleotides and / or the nanoparticles may be functionalized so that the oligonucleotides can be a...

example 1

Preparation of Oligonucleotide-Modified Gold Nanoparticles

A. Preparation of Gold Nanoparticles

[0268] Gold colloids (13 nm diameter) were prepared by reduction of HAuCl.sub.4 with citrate as described in Frens, Nature Phys. Sci., 241, 20 (1973) and Grabar, Anal. Chem., 67, 735 (1995). Briefly, all glassware was cleaned in aqua regia (3 parts HCl, 1 part HNO.sub.3), rinsed with Nanopure H.sub.2O, then oven dried prior to use. HAuCl.sub.4 and sodium citrate were purchased from Aldrich Chemical Company. Aqueous HAuCl.sub.4 (1 mM, 500 mL) was brought to reflux while stirring. Then, 38.8 mM sodium citrate (50 mL) was added quickly. The solution color changed from pale yellow to burgundy, and refluxing was continued for 15 min. After cooling to room temperature, the red solution was filtered through a Micron Separations Inc. 1 micron filter. Au colloids were characterized by UV-vis spectroscopy using a Hewlett Packard 8452A diode array spectrophotometer and by Transmission Electron Microsc...

example 2

Formation of Nanoparticle Aggreates

A. Preparation of Linking Oligonucleotide

[0277] Two (nonthiolated) oligonucleotides were synthesized as described in part B of Example 1. They had the following sequences:

[0278] 3' ATATGCGCGA TCTCAGCAAA [SEQ ID NO:1]; and

[0279] 3' GATCGCGCAT ATCAACGGTA [SEQ ID NO:2].

[0280] Mixing of these two oligonucleotides in a 1 M NaCl, 10 mM phosphate buffered (pH 7.0) solution, resulted in hybridization to form a duplex having a 12-base-pair overlap and two 8-base-pair sticky ends. Each of the sticky ends had a sequence which was complementary to that of one of the oligonucleotides attached to the Au colloids prepared in part C of Example 1.

B. Formation of Nanoparticle Aggregates

[0281] The linking oligonucleotides prepared in part A of this example (0.17 .mu.M final concentration after dilution with NaCl) were added to the nanoparticle-oligonucleotide conjugates prepared in part C of Example 1 (5.1 nM final concentration after dilution with NaCl) at room temp...

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Abstract

The invention provides methods of detecting a nucleic acid. The methods comprise contacting the nucleic acid with one or more types of particles having oligonucleotides attached thereto. In one embodiment of the method, the oligonucleotides are attached to nanoparticles and have sequences complementary to portions of the sequence of the nucleic acid. A detectable change (preferably a color change) is brought about as a result of the hybridization of the oligonucleotides on the nanoparticles to the nucleic acid. The invention also provides compositions and kits comprising particles. The invention further provides methods of synthesizing unique nanoparticle-oligonucleotide conjugates, the conjugates produced by the methods, and methods of using the conjugates. In addition, the invention provides nanomaterials and nanostructures comprising nanoparticles and methods of nanofabrication utilizing nanoparticles. Finally, the invention provides a method of separating a selected nucleic acid from other nucleic acids.

Description

[0001] This application is a continuation-in-part of pending application Ser. No. 09 / 344,667, filed Jun. 25,1999, which was a continuation-in-part of pending application Ser. No. 09 / 240,755, filed Jan. 29, 1999, which was a continuation-in-part of pending PCT application PCT / US97 / 12783, which was filed Jul. 21, 1997, which are incorporated by reference. Benefit of provisional application No. 60 / 031,809, filed Jul. 29, 1996; No. 60 / 200,161, filed Apr. 26, 2000; 60 / 176,409, filed Jan. 13, 2000 is also claimed, the disclosures are incorporated by reference.FIELD OF THE INVENTION[0003] The invention relates to methods of detecting nucleic acids, whether natural or synthetic, and whether modified or unmodified. The invention also relates to materials for detecting nucleic acids and methods of making those materials. The invention further relates to methods of nanofabrication. Finally, the invention relates to methods of separating a selected nucleic acid from other nucleic acids.BACKGROU...

Claims

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Application Information

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Patent Type & Authority Applications(United States)
IPC IPC(8): G01N33/53B82B1/00C07B61/00C07H21/00C12M1/00C12N15/09C12Q1/68C40B40/06G01N21/78G01N27/04G01N33/483G01N33/566G01N33/58G01N37/00
CPCB01J2219/00274Y10S977/924B01J2219/00585B01J2219/00596B01J2219/00603B01J2219/00648B01J2219/00653B01J2219/00659B01J2219/00702B01J2219/00722B82B1/00B82Y15/00B82Y30/00C07B2200/11C07H21/00C12Q1/6816C12Q1/6818C12Q1/6825C12Q1/6827C12Q1/6834C12Q1/6837C12Q1/6839C40B40/00C40B40/06B01J2219/005C12Q2563/155C12Q2563/143C12Q2537/143C12Q2525/197C12Q2525/161C12Q2565/625C12Q2537/125
Inventor MIRKIN, CHAD A.LETSINGER, ROBERT L.MUCIC, ROBERT C.STORHOFF, JAMES J.ELGHANIAN, ROBERTTATON, THOMAS A.GARIMELLA, VISWANADHAMLI, ZHI
Owner NORTHWESTERN UNIV
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